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1.
探讨东方田鼠和小鼠感染日本血吸虫前后不同时点组织与血清中蛋白质电泳谱的变化。分别提取东方田鼠感染日本血吸虫(0、7、12、25 d) 和小鼠感染日本血吸虫(0、7、12、45 d)后的肝脏、脾脏、肺脏蛋白及血清进行聚丙烯酰胺凝胶电泳,对结果比较分析并进行组织病理形态学观察。结果显示,小鼠感染日本血吸虫第7和12天后肝脏150 ku附近蛋白质条带较东方田鼠表达量明显增高;东方田鼠在感染日本血吸虫后40~50 ku附近蛋白质条带较正常东方田鼠表达明显上调;感染第7天后肺脏40 ku附近蛋白质条带较其他时间点变化明显;小鼠感染日本血吸虫第7、12和45天后血清中IgG类蛋白质表达量较东方田鼠显著增高。东方田鼠和小鼠在日本血吸虫感染后不同时点间的组织及血清蛋白组分存在差异,东方田鼠肝脏和血清中150 ku附近蛋白条带较其他动物特异,蛋白质表达丰度高,该蛋白可能与东方田鼠抗日本血吸虫相关。  相似文献   

2.
东方田鼠重复感染日本血吸虫试验研究初步   总被引:11,自引:0,他引:11  
本试验以1000条日本血吸虫尾蚴同时攻击感染东方田鼠洞庭湖种群和东方田鼠宁夏指名亚种进行重复感染对比试验,并在攻击后不同天数分别同时解剖相数目的两种群东方田鼠,收集其体内不同时段,不同部位的日本血吸虫童虫,比较结果证实东方田鼠对日本血吸虫具有天然的抗性,其抗性是可遗传的,不以东方田鼠的地域分布不同、生活环境的差异或传代数目的变化而有所不同,与获得必免疫无关。  相似文献   

3.
本试验以兔、牛、人及感染日本血吸虫后小鼠血清蛋白为对照,分别提取东方田鼠和小鼠的心脏、肝脏、脾脏、肺脏、肾脏、肌肉、皮肤、骨髓和脑组织蛋白,进行聚丙烯酰胺凝胶电泳,综合比较和筛选东方田鼠组织及血清中潜在的抗日本血吸虫相关蛋白组分。结果表明,随着感染日龄的增加,小鼠在感染日本血吸虫后12和45 d,其血清中85 ku附近蛋白质条带较东方田鼠表达量明显上调,且有继续升高的趋势;东方田鼠肝脏及血清中150 ku组分蛋白电泳条带与其他动物相同组织中相应蛋白质在表达量存在显著差异,蛋白质表达丰度高,该蛋白质组分可能与东方田鼠抗日本血吸虫作用相关。  相似文献   

4.
东方田鼠具有天然抗日本血吸虫病特征,为了探索东方田鼠抗日本血吸虫机制,本研究分析了东方田鼠和小鼠感染日本血吸虫尾蚴前后血液细胞以及生化指标的动态变化。东方田鼠和BALB/c鼠定量感染日本血吸虫尾蚴,于感染后第0、1、3、5、8、12、16、21、28、35、42 d分别采集抗凝血和血清,进行血常规和血清生化分析。血常规检测结果表明,东方田鼠感染后第3~16 d,WBC数量显著升高(Neu数量显著升高),而小鼠血常规检测结果正常;血清生化结果显示,东方田鼠感染日本血吸虫后ALB、HDL-C显著升高,小鼠各指标变化不明显。本研究结果显示东方田鼠天然免疫细胞Neu和血清ALB可能与其抗日本血吸虫特性相关,对补体杀伤日本血吸虫机制研究具有重要的指导意义。  相似文献   

5.
把编码中国大陆株日本血吸虫23kD抗原大亲水区多肽的DNA片段亚克隆到pGEX载体上进行表达,并用重组抗原和载体pGEX表达蛋白分别免疫了二批BALB/c小鼠,结果与未免疫小鼠和载体pGEX表达蛋白免疫小鼠相比,重组抗原免疫小鼠分别获得了57.80% ̄70.30%和52.63% ̄62.96%的减虫率,证明重组的日本血吸虫23kD抗原大亲水区多肽可诱导宿主抗血吸虫攻击感染的部分保护力。  相似文献   

6.
IgM单抗体内,外杀伤日本血吸虫的研究   总被引:3,自引:0,他引:3  
本文报道应用急性感染日本血吸虫的Ba1b/c小鼠脾细胞和SP2/0骨髓瘤细胞融合,获得了一株具有抗血吸虫攻击感染保护作用的IgM单抗ssj14。在被动免疫转移试验中,这株单抗在小鼠体内提供了27.69%~68.5%的减虫率。体外ADCC试验显示该单抗参与了巨噬细胞和嗜酸性粒细胞介导的杀伤血吸虫童虫的作用,免疫化学特性测定表明该单抗同时对血吸虫尾蚴、成虫和虫卵三期虫体抗原起反应,靶抗原的分子量为290KD,分别位于血吸虫童虫和成虫的表膜及虫卵的卵壳。靶抗原表位的化学性质为多糖类,它同时被辐照致弱尾蚴免疫的大鼠血清、血吸虫慢性感染人血清和小鼠血清所识别。本文结果表明,IgM抗体和多糖类抗原在血吸虫病免疫保护作用中发挥了重要的作用。  相似文献   

7.
东方田鼠抗日本血吸虫抗体水平检测与分析   总被引:2,自引:0,他引:2  
东方田鼠具有抗日本血吸虫病的特性,观察东方田鼠感染日本血吸虫尾蚴后的特异性抗体水平变化,可为阐明其抗病机制提供依据。本试验用间接ELISA方法检测东方田鼠抗成虫可溶性抗原(SWAP)和几种日本血吸虫基因重组抗原(Sj28-GST、LHD-SjTP1、C-SjParamyosion)的特异性抗体水平动态变化,并检测东方田鼠感染日本血吸虫尾蚴前、后的IgG3亚类特异性抗体水平。结果东方田鼠在攻击日本血吸虫尾蚴后,抗SWAP的特异性抗体水平有所升高,而抗几体基因重组抗原的特异性抗体水平低,没有显著变化。东方田鼠正常血清中抗SWAP和IgG3亚类抗体的OD值是牛血清白蛋白(BSA)对照的31倍。结果表明,在东方田鼠血清中有较高水平的SWAP的IgG3亚类抗体,值得进一步深入探讨。  相似文献   

8.
为了进一步从细胞水平深入研究东方田鼠抗日本血吸虫机制提供试验材料,以及经济可靠地保存野生东方田鼠的生物活性细胞,试验采用胰蛋白酶消化法和组织块贴壁法,分别取妊娠12 d的东方田鼠和昆明小鼠胚胎组织,体外分离、培养成纤维细胞,观察比较不同培养方法、胰蛋白酶浓度、消化时间、传代次数及冻存复苏等因素对分离和培养两种胚胎成纤维细胞的影响,实时观察比较两种细胞生长特性。结果表明:组织块贴壁法和胰蛋白酶消化法均能在体外条件下较好地分离、培养两种胚胎成纤维细胞;采用0.125%胰蛋白酶37℃消化组织10 min,分离的细胞普遍密度大、活力好、贴壁细胞多、速度快,是实验室分离12 d胎龄东方田鼠与小鼠胚胎组织最佳胰酶浓度;昆明小鼠胚胎成纤维细胞较东方田鼠胚胎成纤维细胞足丝长而明显,细胞核较小;两种细胞冻存后生长及活率无显著差异;在含10%血清浓度培养体系中,东方田鼠胚胎成纤维细胞生长速率慢于昆明小鼠胚胎成纤维细胞,生理代数分别为8代和7代,其中2~5代增殖旺盛。说明东方田鼠和昆明小鼠胚胎成纤维细胞在体外分离方法上差异不显著,而两者在生物学特性方面存在一定种属差异。  相似文献   

9.
东方田鼠是迄今为止发现的唯一一种具有天然抗日本血吸虫病特性的哺乳动物,但具体抗病机制尚不明确。本研究应用组织培养法比较了东方田鼠和BALB/c鼠感染日本血吸虫尾蚴后1、3、8、12、15 d的皮肤、肺脏和肝脏中童虫回收情况,结果表明东方田鼠对日本血吸虫的杀伤主要发生在感染后1~3 d和8~12 d。应用RT-PCR和ELISA检测东方田鼠和BALB/c鼠的补体相关分子C3、C9、CD59和DAF的转录水平,结果显示BALB/c小鼠各指标变化不明显,东方田鼠感染后8 d各因子转录水平显著升高。体外杀虫试验结果显示,东方田鼠正常血清和灭活补体血清孵育的日本血吸虫童虫死亡率分别为85.50%±7.15%和60.99%±8.61%,差异具有显著统计学意义(P 0.001)。结果表明东方田鼠补体对日本血吸虫童虫具有显著杀伤效果,东方田鼠早期对日本血吸虫的杀伤作用可能和补体相关。  相似文献   

10.
采用肉眼观察法测定有关药物对日本血吸虫运动性的影响。结果表明:(1)10-3,10-4,10-5mol/Lγ-氨基丁酸(GABA)均有兴奋虫体的作用,其兴奋性随浓度递增而增强;(2)用10-3、10-4mol/L巴氯酚(baclofen)、毕扣扣林(bicuculine)和印防己毒素(picrotoxin)作用虫体后,巴氯酚、毕扣扣林在浓度为10-3mol/L时对虫体有明显抑制作用,浓度为10-4mol/L时抑制作用不明显;而10-3、10-4mol/L印防己毒素均表现一定的兴奋作用;(3)先用10-3mol/LGABA兴奋虫体后,再加入上述药物,结果仍是相同。表明一定浓度的巴氯酚,毕扣扣林不仅对内源性的GABA有拮抗作用,而且对外源性的GABA有拮抗性;而印防己毒素不仅无拮抗性,还有协同GABA的兴奋作用。初步判定,GABA为日本血吸虫的一种兴奋性神经递质,有别于其它寄生虫(如曼氏血吸虫和鸡蛔虫)。  相似文献   

11.
目的寻找与东方田鼠抗性相关的血吸虫靶基因。方法以东方田鼠肺脏裂解物为探针,亲和淘洗筛选日本血吸虫成虫(44d)噬菌体展示cDNA文库。三轮筛选后.随机挑取76个阳性噬菌体克隆进行序列测定及生物信息学分析。结果共获得了13个有效EST序列,其中8条EST序列与已知基因或表达序列标签同源,5个EST序列与已知基因或表达序列标签均无同源性,为新的表达序列标签。功能预测结果显示.这13个有效EST编码蛋白主要与血吸虫基因表达调控及蛋白质加工修饰相关。结论东方田鼠直接或间接地影响血吸虫基因表达,进而影响血吸虫的发育,可能是东方田鼠具有抗血吸虫特性的一个重要分子机理。  相似文献   

12.
Serine proteinase inhibitors (serpins) represent an important superfamily of endogenous inhibitors that regulate proteolytic events active in a variety of physiological functions. Immunological screening of a Schistosoma japonicum adult worm cDNA expression library with sera of Microtus fortis, a naturally resistant vertebrate host, has identified one clone that encoded for a sequence homologous to those of the serpin superfamily. The full-length sequence encoding S. japonicum serpin (Sj serpin) was amplified from adult worm cDNA by using 5'-RACE-PCR and subsequently cloned into the prokaryotic expression vector pET28c. The full-length Sj serpin fusion-protein with his-tag was expressed in E. coli, purified by affinity chromatography and used to immunize New Zealand white rabbits. Sj serpin is located on the tegument in S. japonicum adult worms. C57BL/6 mice immunized with Sj serpin induced the production of high levels of specific IgE and IgG1 subclass antibodies as well as a marked IL-4 response. Lymphocyte surface marker analysis revealed proliferation of CD19 expressing B cells, indicating a predominant Th2-type response to Sj serpin. Immunized mice developed moderate protection against infection of S. japonicum as demonstrated by a 36 and 39% reduction in the recovery of adult worms and eggs, respectively. These data suggested a role for Sj serpin as a vaccine candidate or as a novel target for anti-schistosome drugs.  相似文献   

13.
Fenbendazole in ground feed was fed on 3 successive days at the rate of 3 mg/kg of body weight to 15 sows naturally infected with kidney worm (Stephanurus dentatus). Fifteen similar sows were used as nontreated controls. A total of 49 kidney worms were recovered from daily urine samples collected from 10 of the 15 treated sows within 5 days after the 3rd dose of treatment. One control sow passed 10 kidney worms on the 2nd and 3rd days. Urine samples from treated sows became negative for kidney worm eggs by the 5th to 12th posttreatment days, except for urine samples from 1 sow that contained a few eggs on the 12th day and another sow that contained a few eggs on the 19th and 33rd posttreatment days. Urine samples from control sows contained approximately the same number of kidney worm eggs in the posttreatment period as earlier. Compared with that in control sows, the hatchability of the parasite eggs from treated sows was greatly reduced. Kidney worms were not recovered at necropsy from the treated sows and a total of 860 kidney worms were recovered from the control sows (57 av).  相似文献   

14.
Golden hamsters (Mesocricetus auratus) and Balb C mice were experimentally infected with Trichinella zimbabwensis to determine the effect of host age in the distribution of adult stages in the small intestines. The hamsters and mice were divided into two groups of young and old animals. Hamsters aged 90 days were designated as young and those aged 360 days were designated as old while mice of 30 days of age were designated as young and those aged 90 days as old. To recover the adult parasites of T. zimbabwensis, the small intestines of each animal were separated and divided into four equal parts and each part was slit open longitudinally. The contents were incubated in 0.85% saline for 4 h at 37 degrees C before the adult worms were recovered from the saline. They were fixed in 70% alcohol and counted under a dissecting microscope. In both young and old hamsters and mice, T. zimbabwensis adult worm counts were significantly higher (P < 0.05) in the second segment of the intestines thus invariably reflecting a significantly high count (P < 0.05) in the first (anterior) half of the small intestines. From this study it was demonstrated that host-age had no effect on the distribution of T. zimbabwensis adult worms in the different segments of the small intestines of golden hamsters and Balb C mice.  相似文献   

15.
Ivermectin in solution was given subcutaneously (in the neck) at the rate of 0.5 mg/kg of body weight to 5 sows naturally infected with kidney worm (Stephanurus dentatus). Six similar sows were used as nontreated controls. A total of 114 kidney worms were recovered from daily urine samples of 4 of the 5 treated sows (the 5th sow's samples were negative) during the first 4 days after treatment. Ten kidney worms were recovered from daily urine samples of 1 control sow on the 2nd and 3rd days of the post-treatment period. Examination of urine samples from treated sows for kidney worm eggs showed a precipitous numerical decrease. Treated animals became negative between the 5th and 12th days after treatment, and remained negative until necropsy on the 61st day. Compared with control sows, hatchability of eggs and survival of larvae from treated sows was reduced by 9% and 7%, respectively. Kidney worms were not recovered at necropsy from the treated sows, and a total of 392 kidney worms were recovered from the control sows (av 65).  相似文献   

16.
The aim of this experiment was to establish the oxidative stress model of immune cells in mice induced by PCV2. Optimal infection titer and time of PCV2 were selected. On the 1st,2nd and 3rd day or 1st, 3rd,5th and 7th day,Kunming mice were treated with 100 PCV2 or 10-1 PCV2 by 3 ways (intraperitoneal injection, intranasal administration and intragastrical administration). 7, 14, 21 d post administration, we killed the mice. Reactive oxygen species (ROS),total glutathione (T-GSH),reduced glutathione (GSH),oxidized glutathione (GSSG) levels and xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthase (iNOS) activity were determined to investigate the relationship between the infection time point and the change of ROS level after PCV2 infection. Thus to establish mice immune cells oxidative stress model. The results showed that infection with 100 PCV2 virus in 3 ways every day both at day 1, 2, 3 and 1 mL per mouse was the best program for the follow-up experiment. The intracellular level of ROS of mice infected with PCV2 was extremely significantly increased both at 7, 14 and 21 d (P < 0.01);7 and 14 d post infection, GSH level of PCV2 infected mice had significant difference compared with that of control group (P < 0.05). 7 d after infection, GSSG level was extremely significantly higher than that of control group (P < 0.01); 7 d after infection, T-GSH level was significantly lower than that of control group (P < 0.05), T-GSH was extremely significantly lower than that of control group at 14 d post infection (P < 0.01). XOD, MPO and iNOS activity between PCV2 infection group and blank control group were significantly different at 7 d post infection (P < 0.05). It suggested that PCV2 successfully infected mice, experimental infection programme was Kunming mice were treated with 100 PCV2 by three ways (intraperitoneal injection, intranasal administration and intragastrical administration) both at the 1st,2nd and 3rd day of the experiment.The best condition to establish the oxidative stress model of immune cells in mice was using 100 PCV2 to infect for 7 days.  相似文献   

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